Modern proteomics tasks necessitate combining fast high-performance chromatographic separations with high-resolution mass spectrometers capable of recording tens of high- quality mass spectra per second. While retaining high mass accuracy of recorded spectra, high throughput can be achieved in modern time-of-flight mass spectrometer (QTOF-MS). Important task is to increase QTOF mass resolution to 50-60,000 to a level comparable to that in Orbitrap for ions with mass-to-charge ratios exceeding 800. The main goal of this proposal is designing and building key components of a fast (at least 20 averaged mass spectra per second) Q-TOF mass spectrometer to regularly achieve mass resolution over 50,000 over 800-10,000 Da mass range and demonstrate dynamic range exceeding 5 orders of magnitude. These include cryogenically cooled ion guide interface for efficient collisional cooling of ions before their injection into the of TOF-MS;tilted liner with two ion mirrors to provide longer flight times;data acquisition system with sub-ppm timing accuracy;and combined (analog and digital) multi-channel signal acquisition system. This new high-resolution TOF instrument will be used to provide quantitative analysis of complex protein digest mixtures containing potential biomarkers and pharmaceutical targets. PUBLIC HEALTH RELEVANCE: The development of instruments for high-throughput mass spectrometric analysis plays an important role in modern proteomics and its clinical applications, as well as in drug discovery. We propose to build a novel very high-resolution time-of-flight mass spectrometer, which will utilized cryogenically cooled subsystems, refined design of the ion optical parts, and multi-channel data acquisition to increase reliability and throughput of mass analysis of complex mixtures of protein digests.